Bottom-Up Assembly of Functional Salivary Gland Tissues

功能性唾液腺组织的自下而上组装

基本信息

批准号：
10546502
负责人：
Xinqiao Jia
金额：
$ 45.81万
依托单位：
UNIVERSITY OF DELAWARE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10546502
关键词：
3-Dimensional Ablation Acetylcholine Acinar Cell Acinus organ component Affect Alkenes Animal Model Architecture Basement membrane Beds Biochemical Biological Markers Biomechanics Biomedical Engineering Blood Vessels Body Weight decreased Calcium Cancer Patient Carbachol Cell Communication Cell Differentiation process Cell Lineage Cell Polarity Cells Cellular Spheroids Chemicals Coculture Techniques Complex Cues Deglutition Dental Development Duct (organ) structure Ductal Epithelial Cell Eating Encapsulated Endothelial Cells Endothelium Engineering Environment Epithelium Exhibits Extracellular Matrix Fostering Gel Gene Expression Profile Geometry Gland Goals Growth Growth Factor Head and Neck Cancer Heterogeneity Homeostasis Human Human Engineering Hydrogels Immunocompetent Implant In Vitro Intraperitoneal Injections Investigation Ligation Maintenance Measures Mediating Mesenchymal Mesenchymal Stem Cells Methods Morphogenesis Morphology Myoepithelial cell Natural regeneration Nerve Neurons Neurotransmitters Nude Rats Oral Oral cavity Organ Outcome Measure Parotid Gland Patients Peptide Hydrolases Peptides Phenotype Pilocarpine Property Proteomics Quality of life Radiation therapy Rattus Reaction Resected Saliva Salivary Salivary Gland Tissue Salivary Glands Salivary duct structure Sepharose Shapes Signal Transduction Structure Symptoms Tissue Engineering Tissue Expansion Tissues Treatment Efficacy Trees Vascularization Work Xerostomia alternative treatment analog biomaterial compatibility cell motility cycloaddition head and neck cancer patient hydrogel scaffold implantation in vivo innovation interfacial interstitial mechanical signal mimetics morphogens nerve supply neural neurotrophic factor neurturin polarized cell progenitor reconstitution recruit regeneration potential response restoration side effect stem stem cells subcutaneous transcriptome transcriptome sequencing transplant model treatment strategy

项目摘要

Project Summary Despite advances in treatment strategies, xerostomia (or dry mouth) remains a permanent and devastating side effect of radiotherapy for head and neck cancers, reducing the quality of life for ~50,000 cancer patients each year in the U.S. We aim to develop tissue-engineering approaches to restore salivary function. We have isolated human salivary gland stem/progenitor cells (hS/PCs) from patients prior to radiotherapy. We have created tunable hydrogel matrices that maintain the progenitor status, induce lineage-specific differentiation and promote the development of organized multicellular spheroids from dispersed hS/PCs. Separately, we have engineered salivary gland microtissues that exhibit coordinated calcium activation between hS/PC-derived acini-like core and the surrounding myoepithelial cells. However, a functional gland with extensive branching, polarized acini, and interconnected ducts has not yet been realized. Here, we propose a bottom-up approach to establish functional salivary glands using multicellular assemblies of defined shape, geometry and composition. We will synthesize hydrogel scaffolds that recapitulate key features of the basement membrane and the interstitial matrix in the developing organ. We will reconstitute the vascular, neural and mesenchymal components in the engineered environment to foster tissue morphogenesis in vitro and to maintain tissue homeostasis in vivo. In Aim 1, we will exploit tetrazine ligation, the bioorthogonal and highly efficient cycloaddition reaction between s- tetrazine and strained alkenes, for the establishment of cell-instructive matrices. We will adapt our established methods to generate microgels containing sequestered acetylcholine analog, carbachol (CCh). In Aim 2, we will employ non-adhesive hydrogel microwells to produce multicellular epithelial assemblies consisting of hS/PCs and CCh depots. The resultant microtissue will be encased in a synthetic basement membrane with bioactive peptides to stimulate the development of proacrinar progenitor phenotype. We will generate endothelial microtissues consisting of a core of human salivary gland endothelial cells (hSECs) and a shell of human mesenchymal stem cells (hMSCs). We will co-culture the epithelial and endothelial microtissues in a synthetic extracellular matrix with defined cell-guidance cues to aid in the establishment of a hierarchically integrated tissue assembly. In Aim 3, the engineered gland with integrated microvasculature and conjugated neurotrophic factor, neurturin, will be implanted in the resected parotid bed of athymic rats. Enzymatically triggered release of neurturin will promote implant innervation. Tissue ultrastructure, biomarker expression, gland morphology, biointegration and function will be assessed under various construct configurations. We will interrogate how the engineered microenvironments stimulate differentiation, trigger polarization and promote branching. The overall hypothesis is that hS/PCs co-cultured with hSECs/hMSCs in 3D synthetic matrices displaying biochemical, geometrical and mechanical cues identified from the native organs will assemble into functional salivary tissues. Our investigations will help define bioengineering approaches toward the management of xerostomia.

项目摘要尽管治疗策略取得了进展放射疗法对头颈癌的影响，降低了约50,000名癌症患者的生活质量在美国，我们旨在开发组织工程方法来恢复唾液功能。我们已经孤立了放射疗法之前，患者的人唾液腺茎/祖细胞（HS/PC）。我们创建了可调水凝胶矩阵，可维持祖细胞状态，诱导谱系特异性分化并促进分散的HS/PC的有组织的多细胞球体的发展。单独，我们已经设计了在HS/PC衍生的acini样核心之间表现出协调的钙激活的唾液腺微动物以及周围的肌上皮细胞。但是，具有广泛分支，极化acini的功能腺相互联系的管道尚未实现。在这里，我们提出了一种自下而上的方法来建立使用定义形状，几何形状和组成的多细胞组件的功能性唾液腺。我们将合成水凝胶支架，概括了地下膜和间质基质的关键特征在发育中的器官中。我们将重建该血管，神经和间充质成分设计的环境可在体外促进组织形态发生，并在体内维持组织稳态。在 AIM 1，我们将利用四嗪连接，S-之间的生物正交和高效的环加成反应四嗪和紧张的烯烃，用于建立细胞教学矩阵。我们将适应我们的既定产生含有隔离的乙酰胆碱类似物Carbachol（CCH）的微凝胶的方法。在AIM 2中，我们将采用非粘附水凝胶微孔生产由HS/PC组成的多细胞上皮组件和CCH仓库。最终的微动物将被包裹在具有生物活性的合成基底膜中肽以刺激前骨祖细胞表型的发展。我们将产生内皮由人类唾液腺内皮细胞（HSEC）和人类外壳组成的核心小动物间充质干细胞（HMSC）。我们将在合成中共同培养上皮和内皮微动物细胞外基质具有定义的细胞指示提示，以帮助建立层次结构组织组件。在AIM 3中，具有整体微脉和共轭神经营养的工程腺因素，神经蛋白，将植入切除的无胸腺大鼠的腮腺床中。酶触发的释放神经蛋白会促进植入神经。组织超微结构，生物标志物表达，腺形态，生物整合和功能将在各种构造配置下进行评估。我们将询问如何设计的微环境刺激分化，触发极化并促进分支。总体假设是与HSEC/HMSC共同培养的3D合成矩阵，显示生化，从天然器官鉴定出的几何和机械提示将组装成功能性唾液组织。我们的调查将有助于定义对静态管理的生物工程方法。